KR20220043706A - Method for preparing hydrolyzate of Tenebrio molitor protein using NURUK and protease, and hydrolyzate of Tenebrio molitor protein with enhanced antioxidant content - Google Patents

Abstract

The present invention relates to a method for manufacturing a hydrolysate of Tenebrio molitor (mealworm) protein using malt and a protease, and a Tenebrio molitor protein hydrolysate with an increased antioxidant content. According to the present invention, the method can recover a Tenebrio molitor protein hydrolysate in high yield in comparison with treatment using only a protease, and provides a Tenebrio molitor protein hydrolysate with an increased content of antioxidant functional ingredients such as water-soluble peptides and amino acids, thereby providing a useful effect of providing an improved antioxidant functional health functional food composition. The method comprises a hydrolysis step and a hydrolysate recovery step.

Description

TECHNICAL FIELD [0002] Method for preparing hydrolyzate of Tenebrio molitor protein using NURUK and protease, and hydrolyzate of Tenebrio molitor protein with enhanced antioxidant content}

The present invention relates to a method for producing a protein hydrolyzate of Tenebrio molitor using yeast and proteolytic enzyme, and to a hydrolyzate of brown mealworm protein with enhanced antioxidant content prepared therefrom.

The market size of edible insects is expected to increase from 11 trillion won in 2007 to 38 trillion won in 2020. Korea also started paying attention to the insect industry, and after the government announced the five-year comprehensive plan for promoting the insect industry in 2011, it is fostering the insect industry through policy and deregulation. selected as one of the tasks. The domestic market size grew from 157 billion won in 2009 to 200 billion won in 2014, and is expected to grow to 700 billion won in 2020.

The brown mealworm ( Tenebrio molitor ) is an insect of the Coleoptera family, also called "brown mealworm", and is currently distributed all over the world, including Korea, has strong adaptability, has a short metamorphosis period, and is easy to breed, so it has the advantages of easy industrialization. , was registered as an edible insect raw material with a high protein material in the Food Fair in 2016.

Mealworm brown is a highly versatile raw material as a high-protein material, and it is known that the content of antioxidant functional ingredients such as water-soluble peptides and amino acids in the hydrolyzate of brown mealworm protein is increased. In one study, an attempt was made to develop a food material by treating brown mealworm with a commercial proteolytic enzyme to enhance the antioxidant functional component content of the protein hydrolyzate of brown mealworm (Patent Publication No. 10-2018-0010067). However, the single treatment of commercial proteolytic enzymes hydrolyzes proteins with specificity due to substrate specificity. Even if the amount of proteolytic enzyme treatment is increased, the yield of the hydrolyzate cannot be increased more than a certain level, rather causing problems such as off-flavor. Therefore, only the treatment of commercial proteolytic enzymes alone does not sufficiently derive the antioxidant functional ingredients of brown mealworm protein.

Under this background, the present inventors confirmed that the hydrolyzate from the brown mealworm protein raw material can be recovered in a higher yield than the commercial proteolytic enzyme alone treatment by complex treatment of yeast, a traditional Korean alcoholic beverage fermenting agent, and a commercial proteolytic enzyme. and also confirmed that a hydrolyzate of brown mealworm protein with enhanced antioxidant functional component content can be provided, thereby completing the present invention.

An object of the present invention is to solve the problem of low yield of protein hydrolyzate, which could not be increased more than a certain level with commercial proteolytic enzyme treatment alone, and the problem of not sufficiently extracting the antioxidant functional ingredients of brown mealworm protein. To provide a method for preparing a decomposition product.

Another object of the present invention is to provide a protein hydrolyzate of brown mealworm with enhanced antioxidant functional component content.

In order to achieve the above object,

The present invention, brown mealworm ( Tenebrio molitor ) by treating yeast and proteolytic enzymes to hydrolyze the brown mealworm protein; and

It provides a method for producing a protein hydrolyzate of brown mealworm, comprising; recovering a protein hydrolyzate from the yeast and proteinase-treated brown mealworm.

In addition, the present invention provides a hydrolyzate of brown mealworm protein, prepared by the above production method.

Furthermore, the present invention provides a health functional food composition for antioxidants containing the hydrolyzate of the brown mealworm protein.

In the method for producing a protein hydrolyzate of brown mealworm of the present invention, it is possible to recover the protein hydrolyzate from brown mealworm in a high yield by complex treatment of yeast and proteolytic enzyme, and it is also possible to increase the content of antioxidant functional ingredients, the present invention The hydrolyzate of brown mealworm protein prepared by the method of

1 shows a hydrolysis process diagram for brown mealworm raw material.
Figure 2 shows the measurement of the free amino acid content of the protein hydrolyzate of brown mealworm according to the conditions of treatment with proteolytic enzyme alone, yeast alone treatment, or yeast and proteolytic enzyme complex treatment.
3 is a view confirming the molecular weight composition range of the protein hydrolyzate of brown mealworm according to the conditions of protein raw material and proteinase alone, yeast alone treatment, or yeast and proteinase complex treatment.
4 shows the ABTS antioxidant activity test results of brown mealworm protein hydrolyzate according to the conditions of treatment with proteolytic enzyme alone, yeast alone treatment, or yeast and proteinase complex treatment, and peptides of 3 kDa or less or more than 3 kDa Antioxidant activity of each fraction is shown.
5 shows the ABTS antioxidant activity test results of brown mealworm protein hydrolyzate according to the conditions of treatment with proteolytic enzyme alone, yeast alone treatment, or yeast and proteinase complex treatment, 10 kDa or less, 10-30 kDa or Antioxidant activity of each peptide fraction exceeding 30 kDa is shown.

Hereinafter, the present invention will be described in detail.

The present invention comprises the steps of hydrolyzing brown mealworm protein by treating yeast and proteolytic enzymes on brown mealworm; and

It provides a method for producing a protein hydrolyzate of brown mealworm, comprising; recovering a protein hydrolyzate from the yeast and proteinase-treated brown mealworm.

Conventional commercial proteolytic enzyme treatment alone mainly hydrolyzes proteins with specificity due to the substrate specificity of the proteolytic enzyme, and the hydrolysis effect is insufficient for complex proteins such as brown mealworm protein. There is a problem, and even if the throughput is increased, the time of the hydrolysis process is shortened, and the yield cannot be increased more than a certain amount. In addition, commercial proteolytic enzyme treatment alone has a problem in that it cannot sufficiently extract various water-soluble peptides and amino acid components of brown mealworm protein due to substrate specificity.

The method for producing a protein hydrolyzate of brown mealworm of the present invention is a method for producing a protein hydrolyzate of brown mealworm by complex treatment of yeast, a traditional Korean alcohol fermenting agent, and a commercial proteolytic enzyme. From the action of various proteolytic enzymes, it is possible to hydrolyze even proteins for which commercial proteolytic enzymes do not have specificity. The yield can be increased, and the type and content of water-soluble peptides and amino acids extracted from brown mealworm protein raw materials can be increased.

In one aspect of the present invention, the yeast (NURUK) is a traditional liquor fermentation agent in Korea, which is made by propagating yeast mold by floating grains, and the yeast mold is, for example, Aspergillus oryzae, Heukgukgyun (Aspergillus niger). , may be obtained by using any one or more of Hong Guk-gyun (Monascus anka, Monascus purpureus, Monascus barkeri) and Baek Guk-gyun (Aspergillus luchuensis). On the other hand, the yeast includes commercially available yeast in addition to yeast produced by a traditional method, and improved yeast can also be used.

In another aspect of the present invention, the proteolytic enzyme is an enzyme having hydrolytic activity for brown mealworm protein, and the proteolytic enzyme is, for example, Flavorzyme, Protamax, It is at least one selected from the group consisting of Alcalase and Neutrase. In addition, the treatment of the hydrolase can be treated by combining Flavorzyme, Protamax, Alcalase and Neutrase, for example, Neutrase, Al It can be treated with Kalease, Protamax, and Flavorzyme, and can be treated with Alkalease and Flaverzyme.

The yeast and proteolytic enzyme may be added in an amount of 0.1 to 20% by weight, for example, 0.2 to 18% by weight, 0.3 to 15% by weight or 0.5 to 10% by weight, based on the weight of the brown mealworm raw material. can be

In the case of the yeast, based on the weight of the brown mealworm raw material, it may be added in an amount of 1 to 20% by weight, for example, 2 to 18% by weight, 3 to 15% by weight or 5 to 15% by weight. , For example, 5 to 15% by weight or 10% by weight of commercial yeast may be used, or 5 to 15% by weight or 10% by weight of improved yeast may be used.

In one embodiment of the present invention, in the case of the proteolytic enzyme, it may be added in an amount of 0.1 to 10% by weight, for example, 0.1 to 8% by weight, 0.1 to 5% by weight, based on the weight of the brown mealworm raw material. %, 0.1 to 3% by weight, 0.2 to 2% by weight or 0.5 to 1.5% by weight, for example 0.5 to 1.5% by weight of Alkalease or 0.5 to 1.5% by weight of Flaverzyme, Alternatively, 0.5 to 1.5% by weight of an enzyme mixed with alkaliase and flavorzyme may be used.

On the other hand, the treatment time of the yeast and proteolytic enzyme is 30 minutes to 30 hours, 40 minutes to 25 hours, 60 minutes to 20 hours, 2 to 20 hours, 5 to 20 hours, 10 to 20 hours, or 15 to 20 hours. hours, 17 to 19 hours, or about 18 hours.

The treatment time of the yeast may be 5 to 20 hours, 10 to 20 hours, or 15 to 20 hours, 17 to 19 hours, or about 18 hours.

The treatment time of the proteolytic enzyme may be 30 minutes to 5 hours, 40 minutes to 3 hours, 60 minutes to 2 hours, 70 minutes to 110 minutes, 80 minutes to 100 minutes, or about 90 minutes.

On the other hand, the pH during the treatment of the yeast and proteolytic enzyme is 4 to 8 or 4.5 to 7.

When the yeast is treated, the pH is 4 to 6, 4.2 to 5, or about 4.5.

When the proteolytic enzyme is treated, the pH is 6 to 8, 6.5 to 7.5, or about 7.

On the other hand, the yeast and proteolytic enzyme treatment when the temperature is 20 to 80 ℃, 30 to 70 ℃, 40 to 60 ℃, 45 to 55 ℃, or about 50 ℃.

The yeast treatment temperature is 20 to 50 ℃, 30 to 40 ℃, 35 to 40 ℃, or about 36 ℃.

The temperature during the treatment of the proteolytic enzyme is 30 to 80 ℃, 40 to 70 ℃, 40 to 60 ℃, or about 50 ℃.

For the step of recovering the protein hydrolyzate of brown mealworm, a known method can be used without limitation, for example, after the hydrolysis reaction, centrifugation is performed to separate the layers, and from taking the supernatant, the hydrolyzate of brown mealworm protein can be recovered. there is.

In the hydrolysis step of brown mealworm protein, the brown mealworm may further include drying, powdering and sterilizing, and it may be hydrolyzed by using it as a powdered mealworm raw material. For the drying, powdering and sterilization steps, any known method may be used without limitation.

After the protein hydrolysis step, an enzyme inactivation step may be further included in order to terminate the proteolytic reaction. In the enzyme inactivation step, a known method can be used without limitation, for example, the activity of the proteolytic enzyme can be inactivated by adjusting the pH or temperature, and in one embodiment of the present invention, the reaction system temperature is 70 to 90 ℃ or about 80 ℃, and treatment for 5 to 30 minutes, 10 to 20 minutes or about 15 minutes to inactivate the enzymatic reaction.

In another aspect of the present invention, the method for producing a protein hydrolyzate of brown mealworm may further include the step of fractionating the recovered protein hydrolyzate of brown mealworm to obtain a fraction. The fraction may be used without limitation as long as it is a known protein fractionation method, for example, it may be fractionated by dividing the recovered brown mealworm protein hydrolyzate according to the molecular weight size. In addition, the fraction obtained from the fractionation step may be a fraction of 30 kDa or less, 20 kDa or less, or 10 kDa or less, or a fraction of 3 to 30 kDa, 3 to 20 kDa, or 3 to 10 kDa.

In one embodiment of the present invention, the present inventors have determined the quality characteristics of each brown mealworm protein hydrolyzate obtained by proteolytic alone treatment, yeast alone treatment, or yeast and proteolytic enzyme complex treatment on brown mealworm protein raw materials As a result of comparison, superior quality characteristics were confirmed in the yeast and proteolytic enzyme complex treatment group compared to the protein hydrolysis alone treatment. Specifically, as a result of confirming the yield of the obtained brown mealworm protein hydrolyzate for each treatment condition, it was confirmed that the yield was improved to 47% in the yeast and proteolytic enzyme complex treatment group compared to the protease alone treatment group (see Table 2) ). In addition, as a result of analyzing the free amino acid content in the hydrolyzate of brown mealworm obtained by each treatment condition, the amino acid content of the protein hydrolyzate of brown mealworm was found to be higher in the yeast and proteinase complex treatment group compared to the proteinase alone treatment group. It was confirmed that the increase (see FIG. 2). In addition, as a result of analyzing the organic matter content of the obtained brown mealworm protein hydrolyzate for each treatment condition, it was confirmed that more than about 77% of the protein component contained in the brown mealworm raw material was extracted through the complex treatment with yeast and proteolytic enzyme (Table) see 3).

In addition, as a result of analyzing the antioxidant activity of the obtained brown mealworm protein hydrolyzate for each treatment condition, superior antioxidant activity was confirmed in the yeast and proteinase complex treatment group compared to the protease alone treatment group. In addition, as a result of analyzing the antioxidant activity by size of the obtained protein hydrolyzate, it was confirmed that the antioxidant activity of low molecular weight peptides of 3 kDa or less was superior to high molecular weight proteins exceeding 3 kDa (see FIG. 4 ), and in the peptide fraction of less than 10 kDa, protein Compared to the antioxidant activity of 65% of the hydrolase alone treatment group, the antioxidant activity of peptides less than 10 kDa was found to be 70% or more in the yeast and proteolytic enzyme complex treatment group, confirming that the content of antioxidant functional components was increased ( see Fig. 5).

In addition, the present invention provides a hydrolyzate of brown mealworm protein, prepared by the above production method.

The hydrolyzate of brown mealworm protein obtained through yeast and proteolytic enzyme treatment of the present invention is a hydrolyzate obtained by extracting at least about 77% of the protein component contained in brown mealworm raw material, and is a hydrolyzate obtained by conventional protease alone treatment. Since the free amino acid content is high compared to the hydrolyzate of the obtained brown mealworm protein, it can be used as a food composition rich in amino acid content, a feed composition, and the like.

Furthermore, the present invention provides a health functional food composition for antioxidants containing the hydrolyzate of the brown mealworm protein.

The hydrolyzate of brown mealworm protein obtained through yeast and proteolytic enzyme treatment of the present invention has a higher content of the peptide fraction having antioxidant activity compared to the hydrolyzate of brown mealworm protein obtained through conventional protease alone treatment. There is provided a health functional food composition with enhanced antioxidant functionality as a bar exhibiting antioxidant activity.

Hereinafter, Examples and Experimental Examples of the present invention will be specifically illustrated and described below. However, the Examples and Experimental Examples to be described below are only illustrative of a part of the present invention, and are not limited thereto.

<Example 1> Preparation of protein hydrolyzate of brown mealworm treated with yeast and proteolytic enzyme

<1-1> Preparation of raw material for brown mealworm

The brown mealworm (Mealworm; Tenebrio molitor L.) larvae used in the present invention produced by an insect breeding farm located in Damyang, Jeollanam-do were fasted for 72 hours, washed and sterilized, dried with hot air, and then pulverized with a household grinder. Brown mealworm raw powder was prepared.

<1-2> Preparation of brown mealworm protein hydrolyzate

After adding an appropriate amount of water to the prepared brown mealworm raw powder, 0.5% by weight of Alcalase and 1% by weight of Flavorzyme based on the weight of the raw powder is added and treated at pH 7 and 50°C for 90 minutes, then commercialized based on the weight of the raw powder A hydrolysis reaction was carried out by adding 10% by weight of yeast and processing at pH 4.5 and 36° C. for 18 hours. After hydrolysis, the layers were separated through centrifugation, and a portion of the supernatant was recovered as a hydrolyzate of brown mealworm protein. The precipitate was divided into an insoluble residue derived from the brown mealworm skin (chitin). The supernatant and precipitate were each lyophilized and sampled was used as

<Comparative Example 1> Preparation of brown mealworm protein hydrolyzate treated with yeast alone

After adding an appropriate amount of water to the brown mealworm raw material powder prepared in <1-1>, 10% by weight of commercial yeast was added based on the raw material powder weight and treated at pH 4.5 and 36° C. for 18 hours to perform a hydrolysis reaction. carried out. After hydrolysis, the layers were separated through centrifugation, and a portion of the supernatant was recovered as a hydrolyzate of brown mealworm protein. The precipitate was divided into an insoluble residue derived from the brown mealworm skin (chitin). The supernatant and precipitate were each lyophilized and sampled was used as

<Comparative Example 2> Preparation of protein hydrolyzate from brown mealworm treated with proteolytic enzyme alone

After adding an appropriate amount of water to the brown mealworm raw powder prepared in <1-1>, 0.5 wt% of Alcalase and 1 wt% of Flavorzyme were added to the raw powder weight and treated at pH 7 and 50°C for 90 minutes. A hydrolysis reaction was performed. After hydrolysis, the layers were separated through centrifugation, and a portion of the supernatant was recovered as a hydrolyzate of brown mealworm protein. The precipitate was divided into an insoluble residue derived from the brown mealworm skin (chitin). The supernatant and precipitate were each lyophilized and sampled was used as

The hydrolysis reaction conditions carried out in Example 1 and Comparative Examples 1 and 2 are shown in Table 1 below.

experimental group Applied enzyme and amount used (wt%) Reaction conditions (pH, temperature, time) Example 1 MW - CN - AF CN(10), A(0.5) F(1) After treatment at pH 7, 50°C, 90 minutes,
pH 4.5, 36°C, 18 hours treatment Comparative Example 1 MW - CN CN(10) pH 4.5, 36°C, 18 hours treatment Comparative Example 2 MW - AF A(0.5) F(1) pH 7, 50°C, 90 min treatment

(In the table above, MW: Mealworm A: Alcalase 2.4L, F: Flavorzyme 100L, C: celluclast, V: viscozyme, CN: Commercial yeast)

<Experimental Example 1> Yield of brown mealworm protein hydrolyzate

Separate the brown mealworm protein hydrolyzate supernatant from the insoluble precipitate obtained after centrifugation in Example 1 and Comparative Examples 1 and 2, accurately measure the weight of each sample recovered after lyophilization, and derive the hydrolysis yield by the following formula and the results are shown in Table 2 below.

experimental group Hydrolysis yield (%) Example 1 MW - CN - AF 47 Comparative Example 1 MW - CN 30.18 Comparative Example 2 MW - AF 41.08

(In the table above, MW: Mealworm A: Alcalase 2.4L, F: Flavorzyme 100L, C: celluclast, V: viscozyme, CN: Commercial yeast)

Referring to Table 2, the hydrolyzate yield of the protease alone treatment group was about 41%, and it is confirmed that the hydrolysis yield of the yeast and proteolytic enzyme complex treatment group was improved to 47%.

<Experimental Example 2> Free amino acid content of brown mealworm protein hydrolyzate

The AccQ-Tag method was used to measure the free amino acid content of the hydrolyzate of brown mealworm protein obtained in Example 1 and Comparative Examples 1 and 2, and Waters AccQ-Tag Ultra Derivatization kit (Waters, Milford, MA) was used for amino acid derivatization. , USA) were used. For the analysis, Waters ACQUITY UPLC system (Waters, USA) and 2.1×100 mm AccQ-Tag Ultra column (Waters, USA) were used, and the mobile phase solvent was AccQ-Tag Ultra Eluent A & B (Waters, USA), and the sample was 20 ℃, column 55 ℃, detector 260 nm, flow rate was measured using the gradient (gradient) presented in the kit under the conditions of 0.7 mL / min. As an amino acid standard, amino acid standard H (Thermo Fisher Scientific Inc., Rockford, IL, USA) was used, and the results are shown in FIG. 2 .

2, it is confirmed that the free amino acid content of the yeast and proteolytic enzyme complex treatment group of the present invention is improved compared to the free amino acid content of the protease alone treatment group.

<Experimental Example 3> Organic content of brown mealworm protein hydrolyzate

In order to analyze the change in the organic element (carbon, nitrogen) content of the brown mealworm protein raw material and the brown mealworm protein hydrolyzate obtained in Example 1 and Comparative Examples 1 and 2, the supernatant and insoluble residues after the hydrolysis reaction were fractionated and lyophilized ( MCFD8518, Ilshin Lab Co. Lat, Seoul, Korea) was used to uniformly grind the freeze-dried sample for 120 hours under the conditions of a condenser temperature of -80°C and a pressure of 5 mmTorr, and used as an analysis sample. Carbon and nitrogen analysis was performed by reducing N ₂ O to N ₂ and measuring and quantifying the volume of N ₂ gas by the Dumas method. After weighing 0.2 g each, an elemental analyzer (Elementary, vario MAX cube., Germany) was used, and crude protein The content was expressed by multiplying the nitrogen concentration by the brown mealworm protein conversion factor of 4.76, and the results are shown in Table 3 below.

experimental group Nitrogen content (%) protein content supernatant insoluble residue supernatant Example 1 MW - CN - AF 7.1441 9.21 34.0 Comparative Example 1 MW - CN 5.2353 10.10 24.9 Comparative Example 2 MW - AF 6.25 10.839 29.7 Raw MW
(brown mealworm protein raw material) 9.27 44.1

For brown mealworm protein, the low molecular weight protein component is dissolved in the aqueous solution component through hydrolysis for each treatment condition, so to check the water-soluble protein content in the sample, the nitrogen content was checked and this was converted into protein content.

Looking at Table 3, it was confirmed that the protein content of 44.1% contained in the raw material of brown mealworm was 29.7% in the hydrolyzate of the protease alone treatment group, and about 67% was recovered, and the yeast and proteolytic enzyme complex of the present invention It is confirmed that the treated group hydrolyzate contains 34.0% and recovers about 77%.

On the other hand, there is a research report that the reason that the nitrogen content in insoluble residues is measured as high as 10% is that chitin, which constitutes the outer wall of edible insects such as brown mealworm, consists of unit substances containing non-protein nitrogen such as glucosamine, Accordingly, it was judged that the content appeared to be high.

<Experimental Example 4> Peptide molecular weight range of brown mealworm protein hydrolyzate

<4-1> Measurement of composition range of water-soluble peptide fraction using FPLC to confirm molecular weight of brown mealworm protein hydrolyzate

The experimental conditions for confirming the molecular weight composition range of the brown mealworm protein raw material and the brown mealworm protein hydrolyzate obtained in Example 1 and Comparative Examples 1 and 2 are as follows. A column for protein size exclusion chromatography (SEC, size exclusive chromatography; Superdex 30 increase 10/300 GL,) was mounted on an FPLC (AKTA explorer 100, GE healthcare, Sweden) analysis device, and the supernatant was Using the same amount of the freeze-dried component, a solution dissolved in the same unit volume was injected at 10 mg/mL, and eluent buffer (phosphate saline buffer) was flowed at a flow rate of 0.5 ml/ml and passed through the column after hydrolysis under each condition. The molecular weight of the water-soluble peptide contained in was analyzed, and the results are shown in FIG. At this time, in the case of the SEC method, since the polymer material passes through the column first, it can be inferred that the material eluted at 10 minutes has a higher polymer form than the material eluted at 60 minutes.

<4-2> Results of molecular weight composition range of water-soluble peptide fraction of brown mealworm protein hydrolyzate

3, it was confirmed that the brown mealworm protein raw material was composed of a water-soluble peptide material that passed between 10-20 minutes and 55-60 minutes, and the material confirmed for 15 minutes in the brown mealworm protein raw material was brown mealworm proteolytic enzyme alone It was confirmed that the hydrolyzate of the treatment group was changed to a substance that eluted between 30-40 minutes. On the other hand, in the hydrolyzate of the yeast alone treatment group, it was confirmed that there were few substances eluted in 10-20 minutes and hydrolyzed into characteristic substances at 38 minutes, 45 minutes, and 56 minutes. In addition, the result of the chromatogram of the hydrolyzate of the yeast and proteolytic enzyme complex treatment group showed that the content of the water-soluble peptide constituting the same amount of the sample was increased, and the peak eluted at 40 minutes, 44 minutes, 58 minutes, and 60 minutes. It could be inferred through the area, and it was confirmed that the effect of the composite treatment result was excellent.

<Experimental Example 5> Antioxidant activity of brown mealworm protein hydrolyzate

<5-1> Measurement of antioxidant activity by fractions of 3 kDa or less or more than 3 kDa of brown mealworm protein hydrolyzate

In order to confirm the antioxidant activity of the protein hydrolyzate of brown mealworm, ABTS antioxidant activity was tested. At this time, in order to confirm the size of the hydrolyzed peptide effective for antioxidant activity, the antioxidant activity was quantified by classifying the peptide into fractions of 3 kDa or less or more than 3 kDa using a centricon type membrane filter.

Total antioxidant activity of brown mealworm decomposition product was shown by the radical scavenging activity of ABTS (2,2-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt). ABTS was reacted with potassium persulfate to prepare radical cations, and then, 1 mL of ABTS solution was reacted with the sample for 6 minutes, and the percentage of radical scavenging ability was calculated and shown in FIG. 3 .

3, it is confirmed that the yeast and proteolytic enzyme complex treatment group of the present invention compared to the protease alone treatment group showed superior antioxidant activity in both peptide fractions of 3 kDa or less or more than 3 kDa.

<5-2> Measurement of antioxidant activity by fractions of 10 kDa or less, 10-30 kDa or more than 30 kDa of brown mealworm protein hydrolyzate

In the case of the antioxidant activity of the peptide exceeding 3 kDa confirmed in <5-1>, it may be a value that appears due to the effect of concentrating a material with a relatively large molecular weight in the fractionation process by molecular weight size of the ultrafiltration method, so it is a protein hydrolyzate of brown mealworm After filtering with a 30 kDa size filter, this was again filtered through a 10 kDa size filter, and the antioxidant activity was analyzed by dividing it into fractions of 10 kDa or less, 10-30 kDa, or more than 30 kDa, and the results are shown in FIG. it was

4, in the case of a peptide fraction of less than 10 kDa, the antioxidant activity of the protease alone treatment group was about 65%, whereas in the yeast and proteolytic enzyme complex treatment group of the present invention, the antioxidant activity of the peptide less than 10 kDa It is confirmed that the activity is about 70% or more.

Claims (10)

Brown mealworm ( Tenebrio molitor ) by treating yeast and proteolytic enzymes to hydrolyze the brown mealworm protein; and
Recovering the protein hydrolyzate from the yeast and proteinase-treated brown mealworm; including, a method for producing a protein hydrolyzate from brown mealworm.
According to claim 1,
The yeast is a method for producing a brown mealworm protein hydrolyzate, characterized in that it is a yeast obtained by using at least one selected from the group consisting of Hwanggukgyun, Heukgukgyun, Honggukgyun, and Baekkukgyun.
According to claim 1,
The short-acting hydrolase is one or more proteolytic enzymes selected from the group consisting of Flavorzyme, Protamax, Alcalase and Neutrase. , Method for producing brown mealworm protein hydrolyzate.
According to claim 1,
The treatment of the proteolytic enzyme is a process for 60 minutes to 120 minutes at a pH of 6 to 8 and a temperature of 40 to 60 ℃, the method for producing a protein hydrolyzate of brown mealworm.
According to claim 1,
The treatment of the yeast is a method for producing a hydrolyzate of brown mealworm protein, which is to be treated for 10 to 20 hours at a pH of 4 to 6 and a temperature of 30 to 40 ℃.
According to claim 1,
The treatment of the proteolytic enzyme is treated with 0.1 to 5% by weight based on the weight of the brown mealworm raw material, and the treatment of the yeast is treated with 1 to 20% by weight relative to the weight of the brown mealworm raw material, characterized in that it is a brown mealworm protein hydrolysis A method for producing a decomposition product.
According to claim 1,
The step of hydrolyzing the brown mealworm protein by treating the yeast and proteolytic enzymes,
1) treating brown mealworm proteolytic enzyme in an amount of 0.1 to 5% by weight based on the weight of brown mealworm raw material, at a pH of 6 to 8 and a temperature of 40 to 60° C. for 60 to 120 minutes; and
2) processing the yeast in the brown mealworm treated with the proteolytic enzyme at 1 to 20% by weight based on the weight of the brown mealworm raw material, at a pH of 4 to 6 and a temperature of 30 to 40 ° C. for 10 to 20 hours; A method for producing a brown mealworm protein hydrolyzate, characterized in that it comprises.
The brown mealworm protein hydrolyzate produced by the method of claim 1.
The protein hydrolyzate according to claim 8, wherein the protein hydrolyzate of mealworms brown is a peptide fraction having a molecular weight of 10 kDa or less.
A health functional food composition for antioxidants containing the protein hydrolyzate of claim 8.

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